Fig. 6: Disulfiram (DSF) is a potent inhibitor of chemokine-mediated FROUNT functions.

a Effect of DSF (25 µM) on CCL2-induced pseudopodia protrusion compared with DMSO control and wortmannin (WM, 25 µM). b Effect of DSF on chemotaxis (red line), and cell viability (black line) (mean ± s.e.m., n = 3). c Translocation of PH-Akt-tHcRed to the plasma membrane as an indicator of PI3K activation in CCR2 + Chinese hamster ovary cells expressing PH-Akt-tHcRed, in the presence or absence of DSF upon stimulation with CCL2. Representative confocal microscopic images of cells stimulated with CCL2 (left) and percentage of cells in which PH-Akt-tHcRed was translocated to the membrane. Arrowheads indicate sites of membrane translocation of PH-Akt-tHcRed. More than five visual fields were averaged. d, e The ALDH inhibitor cyanamide did not block FROUNT-CCR2 interactions or chemotaxis (mean ± s.e.m., duplicates from one of three independent experiments.). f ALDH inhibition activity of DSF and cyanamide evaluated by ALDEFLUOR assay (mean ± s.e.m., n = 6). g–i TAXIScan assay⁵⁷ for migration of THP-1 cells treated with DMSO (control) or DSF (50 μM) towards CCL2. g cell tracks, h velocity, and i directionality. Plots indicate means of 28 cell tracks ± s.e.m. *P < 0.05, ***P < 0.001, ****P < 0.0001 by two-tailed unpaired Student’s t-test as compared to control. Scale bars, 20 μm.